Molecular Pathogenesis of Rickettsioses and Development of Novel Anti-Rickettsial Treatment by Combinatorial Peptide-Based Libraries
Abstract
The purpose of this study is to utilize adaptein libraries coded within pantropic retroviral vectors that confer protection against rickettsial pathogens and to study the molecular pathogenesis of rickettsioses. The following specific aims were proposed: 1) To establish heterogeneous cell populations, with each cell expressing a unique member of a complex combinatorial peptide-based (e.g. adaptein) library and challenge with R. prowazekii, R. rickettsii, and O. tsutsugamushi; 2) To determine the role of NF-kB, cytokines (TNFalpha, IFN-gamma, RANTES), ROS and NO in intracellular killing of rickettsia-infected monolayers containing adapteins; and 3) To characterize signal transduction pathways modulating the cytoskeletal events responsible for the increased vascular permeability. During the first year of this project we were able to construct two dozen libraries encoding combinatorial 6-mer, 12-mer, and 18-mer peptides. We successfully produced these libraries in bacterial cells and transfected two different cell lines with recombinant retroviruses containing the libraries with high efficiency for rickettsial challenges. We have also developed in vitro models of endothelial barrier using rat derived microvascular endothelial cells. Measurements of endothelial permeability using FITC-dextrans in transwell settings and using ECIS have been performed. Elevation of intracellular calcium in infected cell monolayers and activation of calmodulin have also been demonstrated.
Document Details
- Document Type
- Technical Report
- Publication Date
- Feb 01, 2003
- Accession Number
- ADA418019
Entities
People
- David H. Walker
Organizations
- University of Texas Medical Branch